Purging of T-lymphocytes with magnetic affinity colloid.
There is a well-documented correlation between the number of T-lymphocytes in the bone marrow graft and subsequent development of acute graft-versus-host disease after allogeneic bone marrow transplantation. The incidence of acute graft-versus-host disease may be decreased with elimination of mature T-lymphocytes from the bone marrow graft. We have developed an immunomagnetic separation procedure using an avidin-based magnetic affinity cobalt colloid. Bone marrow cells were incubated with a combination of CD2, CD3, CD4, and CD8 monoclonal antibodies. The cells were washed and then incubated with the biotinylated, affinity-purified IgG fraction of goat anti-mouse immunoglobulins followed by an avidin-based magnetic affinity colloid. The cells were then run through a high-magnetic gradient separation column utilizing an external samarium cobalt magnet. The number of residual T-lymphocytes was assessed by limiting dilution analysis of clonogenic T-lymphocytes. This procedure produces an approximately 1.7-log reduction of antibody-reactive cells with 45% recovery of hematopoietic progenitors (granulocyte-macrophage colony-forming cells, GM-CFC). This causes a reduction of T-lymphocytes in the bone marrow graft to approximately 5 x 10(5) cells/kg body weight.
Yau
J C
JC
Department of Hematology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Reading
C L
CL
Thomas
M W
MW
Davaraj
B M
BM
Tindle
S E
SE
Jagannath
S
S
Dicke
K A
KA
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000032:analysis; D001360:Avidin; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation; D002469:Cell Separation / Q000379:methods*; D003037:Cobalt Radioisotopes; D003102:Colloids; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D007202:Indicators and Reagents; D008280:Magnetics; D008854:Microscopy, Electron; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D003037:Cobalt Radioisotopes; D003102:Colloids; D007202:Indicators and Reagents; D001360:Avidin